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Alexa Fluor® 647 anti-human CD326 (EpCAM) Antibody

Pricing & Availability
Clone
9C4 (See other available formats)
Regulatory Status
RUO
Other Names
Ep-CAM, tumor associated calcium signal transducer 1, epithelial cell surface antigen, epithelial glycoprotein 2, EGP2, adenocarcinoma associated antigen, TROP1
Isotype
Mouse IgG2b, κ
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Product Citations
publications
1)9C4_Alx647_071007
Human colon carcinoma cell line (HT29) stainined with 9C4 Alexa Fluor® 647
  • 1)9C4_Alx647_071007
    Human colon carcinoma cell line (HT29) stainined with 9C4 Alexa Fluor® 647
  • 9C4_A647_CD326_Antibody_ICC_011421
    MCF7 breast cancer cell line was stained with 4 µg/mL anti-human CD326 Alexa Fluor® 647 and nuclear counterstained with DAPI. Images were acquired with a TE300 fluorescence microscope with a 20x objective. Data provided by: Er Liu and John Nolan, La Jolla Bioengineering Institute
See Alexa Fluor® 647 spectral data
Cat # Size Price Save
324212 100 tests ¥64,460
Description

CD326 is also known as Ep-CAM, tumor associated calcium signal transducer 1, epithelial cell surface antigen, epithelial glycoprotein 2, EGP2, adenocarcinoma associated antigen, and TROP1. CD326 is a type I transmembrane protein containing six disulfide bridges and one THYRO domain. This cell surface glycosylated 40 kD protein is highly expressed in bone marrow, colon, lung, and most normal epithelial cells and is expressed on carcinomas of gastrointestinal origin. Recently, it has been reported that CD326 expression occurs during the early steps of erythrogenesis. CD326 functions as a homotypic calcium-independent cell adhesion molecule and is believed to be involved in carcinogenesis by its ability to induce genes involved in cellular metabolism and proliferation. CD326 antigen is an immunotherapeutic target for the treatment of human carcinomas.

Product Details
Technical data sheet

Product Details

Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
DU.4475 breast carcinoma
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
ICC - Verified
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
Application Notes

Additional reported applications (for the revelant formats) include: immunofluorescence, immunohistochemistry3, and spatial biology (IBEX)4,5.

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

Application References

(PubMed link indicates BioLegend citation)
  1. Lammers R, et al. 2002. Exp. Hematol. 30:537.
  2. Schultz LD, et al. 2010. P. Natl. Acad. Sci. USA 107:13022. PubMed
  3. Human Protein Atlas http://www.proteinatlas.org/ENSG00000119888/antibody (IHC)
  4. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  5. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Stallcop LE, et al. 2018. Lab Chip. 18:451. PubMed
  2. Liu T, et al. 2019. Oncol Lett. 18:2262. PubMed
  3. Freire‐Pritchett P et al. 2017. eLife. 6 pii: e21926. PubMed
  4. Kitajima S, et al. 2019. Cancer Discov. 9:34. PubMed
  5. Csizmar CM, et al. 2019. J Am Chem Soc. 141:251. PubMed
  6. Qin XY, et al. 2020. Cell Death Dis. 0.504166667. PubMed
  7. Rosenbluth JM, et al. 2020. Nat Commun. 11:1711. PubMed
  8. Sun Q, et al. 2018. J Clin Invest. 128:531. PubMed
  9. Morris EJ, et al. 2020. Cell Rep. 30:3605. PubMed
  10. Somerville TD, et al. 2020. eLife. 9:e53381.. PubMed
  11. Campisi M, et al. 2020. Front Immunol. 1.909722222. PubMed
  12. Meirelles K, et al. 2012. Proc Natl Acad Sci U S A. 109:2358. PubMed
  13. Zhou J, et al. 2011. J Vis Exp. 55: 3322. PubMed
  14. Pham K, et al. 2016. Am J Pathol. 186: 1537-1546. PubMed
  15. Hendricks A, et al. 2020. Cancers (Basel). 12:00. PubMed
  16. Liu X, et al. 2021. iScience. 24(6):102551. PubMed
  17. Ebisudani T, et al. 2021. Cell Reports. 35(10):109218. PubMed
  18. Sun L, et al. 2021. Cancer Cell. 39:1361. PubMed
  19. Robertson N, et al. 2020. J Biol Chem. . PubMed
  20. Robertson N, et al. 2020. J Biol Chem. 295:18436. PubMed
  21. Hendricks A, et al. 2021. Front Oncol. 11:646885. PubMed
  22. Arenas EJ, et al. 2021. Nat Commun. 12:1237. PubMed
  23. Vanslambrouck JM, et al. 2022. Nat Commun. 13:5943. PubMed
  24. Liang J, et al. 2022. J Clin Invest. 132: . PubMed
  25. Martínez-Sabadell A, et al. 2022. Cell Rep. 41:111430. PubMed
  26. Estrada HQ, et al. 2022. Inflamm Bowel Dis. 28:667. PubMed
  27. Bhatia S, et al. 2022. Cancer Res. 82:1174. PubMed
  28. Qin XY, et al. 2023. Cell Death Dis. 14:358. PubMed
RRID
AB_756086 (BioLegend Cat. No. 324212)

Antigen Details

Structure
Type I transmembrane protein, contains six disulfide bridges, one THYRO domain, approximate molecular weight 40 kD.
Distribution

Highly expressed in bone marrow, colon, lung, and most normal epithelial cells. Also highly expressed on carcinomas of gastrointestinal origin. Expressed during early erythrogenesis.

Function
Homotypic calcium-independent cell adhesion. CD326 is believed to be involved in carcinogenesis by its ability to induce genes involved in cellular metabolism and proliferation.
Modification
Glycosylated.
Cell Type
Embryonic Stem Cells, Epithelial cells
Biology Area
Cell Biology, Immunology, Stem Cells
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References

1. Strnad J, et al. 1989. Cancer Res. 49:314.
2. Munz M, et al. 2004. Oncogene 23:5748.
3. Rao CG, et al. 2005. Int. J. Oncol. 27:49.

Gene ID
4072 View all products for this Gene ID
UniProt
View information about CD326 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.

Are other fluorophores compatible with IBEX?

Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.

The same antibody works in one tissue type but not another. What is happening?

Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.

How can I be sure the staining I’m seeing in my tissue is real?

In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.

Go To Top Version: 5    Revision Date: 04/26/2022

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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